Department of Gastroenterology and hepatology, the 2nd Affiliate Hospital of Dalian Medical University, Dalian, Liaoning Province, China.
Biomed Pharmacother. 2012 Apr;66(3):167-72. doi: 10.1016/j.biopha.2011.10.002. Epub 2012 Jan 18.
We have previously demonstrated that chloride intracellular channel 1 (CLIC1) is involved in the lymphatic metastasis of tumors. In this study, a self-designed shRNA sequence of mouse CLIC1 gene was synthesized and inserted into a pGPU6/GFP/Neo plasmid, then stably transfected into mouse hepatic carcinoma cell line Hca-F cells to down-regulate the expression of CLIC1 gene. The levels of expression of CLIC1 mRNA and protein were detected by real-time quantitative polymerase chain reaction (qRT-PCR) and western blot (WB) analysis, respectively. The down-regulation of CLIC1 enhanced proliferative activity, increased the ratio of G2/M and decreased percentage of apoptosis. In addition, the capability of migration and invasion decreased significantly. The results indicate that CLIC1 is a critical factor in the development of lymphatic metastasis.
我们之前已经证明氯离子通道蛋白 1（CLIC1）参与了肿瘤的淋巴转移。在这项研究中，我们合成了一个小鼠 CLIC1 基因的自我设计的 shRNA 序列，并将其插入到 pGPU6/GFP/Neo 质粒中，然后稳定转染到小鼠肝癌细胞系 Hca-F 细胞中，以下调 CLIC1 基因的表达。通过实时定量聚合酶链反应（qRT-PCR）和蛋白质印迹（WB）分析，分别检测 CLIC1 mRNA 和蛋白的表达水平。下调 CLIC1 增强了增殖活性，增加了 G2/M 期的比例，降低了细胞凋亡的比例。此外，迁移和侵袭能力显著下降。这些结果表明 CLIC1 是淋巴转移发展的关键因素。
